title: DNA sequencing in HydroLink matrices: extension of reading ability to greater than 600 nucleotides.
abstract: All the systems for optimizing DNA sequencing published so far have introduced modifications regarding: (i) linearization of band migration via ionic strength gradients or wedge-shaped gels; (ii) automatization of band reading via introduction of fluorescent probes; (iii) direct blotting analysis; (iv) pulsed electric fields and (v) discontinuous buffer systems. In all these systems, DNA sequence reading with an accuracy of ca. 98% rarely exceeds a length of 350 bases. We have chosen, in order to increase the reading ability of a single gel, to manipulate the characteristics of the gel matrix. The Seq-HydroLink gel formation here reported allows optimal reading, from a single gel run, of at least 600 bases. In order to guarantee this reading ability in a single run, the upper and lower ends of the ladder are time-resolved, i.e. the same sample is applied to the gel matrix at three different time intervals. The present system represents an increase of at least 30% in reading ability as compared with any type of polyacrylamide gel formulation so far reported.
authors.affiliation: Department Biomedical Sciences and Technologies, University of Milano.
authors.forename: C
authors.initials: C
authors.lastname: Gelfi
authors.forename: A
authors.initials: A
authors.lastname: Canali
authors.forename: P C
authors.initials: PC
authors.lastname: Righetti
authors.forename: P
authors.initials: P
authors.lastname: Vezzoni
authors.forename: C
authors.initials: C
authors.lastname: Smith
authors.forename: M
authors.initials: M
authors.lastname: Mellon
authors.forename: T
authors.initials: T
authors.lastname: Jain
authors.forename: R
authors.initials: R
authors.lastname: Shorr
mesh_terms: D001435:Bacteriophages / Q000235:genetics; D003432:Cross-Linking Reagents; D004247:DNA / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel; D009710:Nucleotide Mapping
chemical_list: D003432:Cross-Linking Reagents; D004247:DNA